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zRbm14 distributes in cells as puncta and forms liquid droplets in vitro through IDR.a zRbm14a displayed punctate distributions in the cytoplasm and nucleus of zebrafish embryonic cells. Zebrafish embryos immunostained as in Fig. 1c were imaged at high resolution. Arrowheads indicate bright perinuclear zRbm14a puncta. b GFP-zRbm14b displayed similar subcellular localizations as endogenous zRbm14a. Zebrafish embryos were microinjected at the one-cell stage with in vitro-transcribed mRNA (800 pg per embryo) to express GFP-zRbm14b, fixed at approximately 4 hpf, and imaged at high resolution. c PLD prediction for Rbm14 orthologues. The diagrams were generated by using the Prion-like Amino Acid Composition (PLAAC) program (http://plaac.wi.mit.edu)1. Sequences with the PLD probability >0.5 (y-axis) are considered as a PLD. d Diagrams of His-GFP-tagged zRbm14b and its mutants. Detailed mutation sites in zRbm14bIDR18S are indicated in Supplementary Fig. 3a. e Time-dependent droplet growth. Twenty micromolar of His-GFP-zRbm14bIDR containing 1% PEG8000 were incubated at 25 °C for the indicated time and imaged for GFP fluorescence. f Concentration-dependent droplet formation. Ten to 40 μM of purified His-GFP or its tagged proteins containing 1% PEG8000 were shifted from ice to 25 °C for 5 min and imaged. Also see Supplementary Fig. 3b, c. g Image sequences showing fusion processes of two droplets (arrows)
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